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Hirudin insensitive thrombin-stimulated platelet release.

D S Reinhold, H Gershman

    Thrombosis Research
    |February 15, 1985
    PubMed
    Summary
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    Platelet stimulation involves two thrombin complexes. Hirudin sensitivity changes, indicating a conversion from an initial to an activated complex, crucial for understanding platelet activation.

    Area of Science:

    • Biochemistry
    • Hematology
    • Cellular Biology

    Background:

    • Platelet activation by thrombin is a complex process.
    • Two proposed platelet-thrombin complexes: initial (P-T) and activated (P*-T).

    Purpose of the Study:

    • Differentiate between hirudin-sensitive and hirudin-insensitive platelet-thrombin complexes.
    • Investigate the conversion of P-T to P*-T and its effect on thrombin stimulation.

    Main Methods:

    • Measuring radioactive serotonin release from platelets as an indicator of thrombin stimulation.
    • Utilizing hirudin, a thrombin inhibitor, to assess complex sensitivity.
    • Conducting experiments at different preincubation times and temperatures (4°C and 37°C).
    • Comparing results in phosphate buffer versus acetate-Tris buffer.

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    Main Results:

    • At short preincubation times (0-2 min) at 4°C in phosphate buffer, hirudin almost completely inhibited serotonin release upon warming, indicating a hirudin-sensitive complex.
    • With longer preincubation (30 min), hirudin's effectiveness decreased, suggesting the formation of a hirudin-insensitive complex.
    • This transition from hirudin-sensitive to insensitive complex was faster in acetate-Tris buffer.
    • Buffer composition (phosphate presence) and ionic strength influenced hirudin's inhibitory effect, not its inhibition constant.

    Conclusions:

    • Evidence supports the existence of two distinct platelet-thrombin complexes.
    • A slow conversion of the initial P-T complex to an activated P*-T complex occurs, influencing thrombin-induced platelet release.
    • Buffer conditions significantly modulate the kinetics of this complex conversion and hirudin's efficacy.